东南大学学报（医学版）
東南大學學報（醫學版）
동남대학학보（의학판）
JOURNAL OF SOUTHEAST UNIVERSITY(MEDICAL SCIENCE EDITION)
2014年
3期
291-296
,共6页
朱洪艳%陈蓉%姜藻
硃洪豔%陳蓉%薑藻
주홍염%진용%강조
内皮祖细胞%胞嘧啶脱氨酶%肝细胞癌%慢病毒载体%凋亡
內皮祖細胞%胞嘧啶脫氨酶%肝細胞癌%慢病毒載體%凋亡
내피조세포%포밀정탈안매%간세포암%만병독재체%조망
endothelial progenitor cells%cytosine deaminase%hepatocellular carcinoma%lentiviral vectors%apoptosis
目的：研究自杀基因胞嘧啶脱氨酶（CD）基因转染的小鼠骨髓源性内皮祖细胞（EPCs）联合酶前体药物5-氟胞嘧啶（5-FC）对小鼠肝癌细胞株H22的体外抗增殖作用。方法：分离普通小鼠骨髓源性EPCs，培养鉴定，Polybrene技术转染含CD基因的慢病毒载体重组体plenti6．3-EGFP-CD至EPCs，Western blotting检测目的蛋白的表达。利用Transwell小室共培养体系，用转染CD基因的EPCs处理H22细胞。 CCK-8法检测H22细胞增殖变化，流式细胞仪检测细胞凋亡水平。结果：成功转染CD基因至EPCs；CCK-8法检测显示随着5-FC浓度增加，细胞增殖逐渐下降，5-FC浓度达100 mg· L-1时，肿瘤细胞增殖抑制率达到（54．74±5．38）％（P＜0．05），细胞平均凋亡率为（48．71±4．62）％（P＜0．05）。结论：CD基因修饰的EPCs 联合5-FC体外可明显抑制肝癌H22细胞增殖，诱导肿瘤细胞凋亡。
目的：研究自殺基因胞嘧啶脫氨酶（CD）基因轉染的小鼠骨髓源性內皮祖細胞（EPCs）聯閤酶前體藥物5-氟胞嘧啶（5-FC）對小鼠肝癌細胞株H22的體外抗增殖作用。方法：分離普通小鼠骨髓源性EPCs，培養鑒定，Polybrene技術轉染含CD基因的慢病毒載體重組體plenti6．3-EGFP-CD至EPCs，Western blotting檢測目的蛋白的錶達。利用Transwell小室共培養體繫，用轉染CD基因的EPCs處理H22細胞。 CCK-8法檢測H22細胞增殖變化，流式細胞儀檢測細胞凋亡水平。結果：成功轉染CD基因至EPCs；CCK-8法檢測顯示隨著5-FC濃度增加，細胞增殖逐漸下降，5-FC濃度達100 mg· L-1時，腫瘤細胞增殖抑製率達到（54．74±5．38）％（P＜0．05），細胞平均凋亡率為（48．71±4．62）％（P＜0．05）。結論：CD基因脩飾的EPCs 聯閤5-FC體外可明顯抑製肝癌H22細胞增殖，誘導腫瘤細胞凋亡。
목적：연구자살기인포밀정탈안매（CD）기인전염적소서골수원성내피조세포（EPCs）연합매전체약물5-불포밀정（5-FC）대소서간암세포주H22적체외항증식작용。방법：분리보통소서골수원성EPCs，배양감정，Polybrene기술전염함CD기인적만병독재체중조체plenti6．3-EGFP-CD지EPCs，Western blotting검측목적단백적표체。이용Transwell소실공배양체계，용전염CD기인적EPCs처리H22세포。 CCK-8법검측H22세포증식변화，류식세포의검측세포조망수평。결과：성공전염CD기인지EPCs；CCK-8법검측현시수착5-FC농도증가，세포증식축점하강，5-FC농도체100 mg· L-1시，종류세포증식억제솔체도（54．74±5．38）％（P＜0．05），세포평균조망솔위（48．71±4．62）％（P＜0．05）。결론：CD기인수식적EPCs 연합5-FC체외가명현억제간암H22세포증식，유도종류세포조망。
Objective:To investigate the suppression of the proliferation of hepatoma H 22 cells treated through mice marrow-derived endothelial progenitor cells(EPCs) modified by cytosine deaminase (CD) gene and enzyme prodrug 5-fluorocytosine (5-FC) in vitro.Methods: EPCs were isolated from mice bone marrow .After cultured and identified, EPCs were transfected by lentiviral vector carrying CD gene (plenti6.3-EGFP-CD ) using Polybrene technique .Expression of target protein was tested by Western blotting .H22 cells were treated by the EPCs carrying CD gene using Transwell cells co-culture system .CCK-8 method was used to detect the proliferation of H 22 cell. The tumor apoptosis was tested by flow cytometry .Results: The transfection of CD gene to EPCs was detected successfully .CCK-8 assay showed that cell proliferation gradually decreased along with increasing of 5-FC concentration .When 5-FC reached 100 mg· L-1 , the proliferation inhibition of tumor cells was about ( 54 .74 ± 5.38)%(P<0.05) and the average apoptosis rate was (48.71 ±4.62)%(P <0.05).Conclusion: EPCs modified by CD gene with 5-FC can effectively inhibit hepatoma H 22 cells proliferation and induce cells apoptosis in vitro.
